- Open Access
CTGF is a central mediator of tissue remodeling and fibrosis and its inhibition can reverse the process of fibrosis
© Lipson et al; licensee BioMed Central Ltd. 2012
- Published: 6 June 2012
CTGF is a secreted matricellular protein with very complex biology. It has been shown to modulate many signaling pathways leading to cell adhesion and migration, angiogenesis, myofibroblast activation, and extracellular matrix deposition and remodeling, which together lead to tissue remodeling and fibrosis. It has been reported in the literature that inhibition of CTGF expression by siRNA prevents CCl4-induced liver fibrosis and can reverse fibrosis when administered after significant collagen deposition is observed. A monoclonal antibody to CTGF that is currently in clinical development (FG-3019) has demonstrated the ability to reverse vascular stiffening and improve cardiac function in a rat model of diabetic complications. FG-3019 has also exhibited activity in a murine radiation-induced pulmonary fibrosis model. When FG-3019 was administered to mice after a significant radiation-induced increase in lung density could be observed by CT imaging, the density of the lungs was observed to decrease over the period during which the antibody was administered and to remain stable after therapy had ceased. When considered together, these data indicate that inhibition of CTGF can prevent and reverse the process of fibrosis.
- Idiopathic Pulmonary Fibrosis
- Connective Tissue Growth Factor
- Tissue Remodel
- Axial Stiffness
- Matricellular Protein
CTGF and its modulation of cell biology
Connective tissue growth factor (CTGF, CCN2) is a member of a small family of proteins that are characterized by their highly conserved disulfide bonding pattern and having 3-4 domains with homology to other proteins [1, 2]. CTGF has four domains: domain 1 is homologous to IGF-1 binding proteins, domain 2 is homologous to the von Willebrand factor type C repeat, domain 3 is homologous to the thrombospondin type 1 repeat and domain 4 contains a cysteine knot motif that is common to proteins that bind to heparan sulfate proteoglycans (HSPGs). The two N-terminal domains (1 and 2) are linked to the two C-terminal domains (3 and 4) by a peptide that is not homologous to other proteins and is proteolytically labile. The N-half of CTGF is proteolytically stable and can often be observed in biological fluids (plasma, urine) of diseased subjects, but is rarely observed at substantial levels in healthy individuals [3, 4]. The C-half of CTGF appears to be proteolytically labile, and is rarely observed in biological fluids. Similarly whole CTGF is rarely observed at significant concentrations in biological fluids, except in subjects with liver disease [5, 6].
Although it was called CTGF when it was discovered , it does not behave like a traditional growth factor or cytokine since it does not appear to have a unique receptor to which it binds with high affinity to induce signal transduction. It may be more accurate to think of CTGF as a matricellular protein that modulates the interaction of cells with the matrix to modify the cellular phenotype [8–11]. Matricellular proteins such as SPARC, osteopontin and thrombospondins represent a sub-class of extracellular matrix (ECM) proteins that do not provide a structural function, but instead modulate cellular functions and signaling through multiple mechanisms that depend on the cell type and context .
The interaction of CTGF with various molecules is thought to positively or negatively alter the signal transduction pathways in which they participate. The result of this modulation of signaling are changes in cell adhesion and migration, angiogenesis and vascular permeability, differentiation, including myofibroblast formation and activation, extracellular matrix deposition and remodeling, all of which together lead to tissue remodeling and changes in organ structure.
Although the dogma in the CTGF literature suggests that all of the biological responses to CTGF are mediated by the direct interaction of CTGF with cytokines or receptors, the mechanisms by which it modulates cell function are likely to be more complicated. There are at least 5 mechanisms by which CTGF could simultaneously modulate the cellular environment and phenotype: 1) CTGF could act as an extracellular adapter protein by binding to cytokines and helping to present them to their receptors to stimulate response (e.g. TGFβ ) or sequestering them in the matrix and thereby preventing them from stimulating signal transduction (e.g. VEGF ); 2) CTGF could competitively bind to HSPGs, which would displace heparin-binding growth factors or receptor antagonists and thereby alter their local concentration and ability to modulate signaling (data not shown); 3) CTGF can block matrix binding sites or create new matrix binding sites, which would alter matrix signaling, cell adhesion and motility (e.g. [14–16]); 4) CTGF can bind directly to cell surface receptors and stimulate signal transduction (e.g. [16–18]); and 5) CTGF may be taken up into cells via endocytic pathways and act as an intracellular adapter protein to modulate signal transduction pathways in the cytoplasm and/or nucleus . Because CTGF can work through so many mechanisms simultaneously, its biology is very complicated and incompletely understood. In addition, response to CTGF will be context dependent, and will vary with matrix, cytokine environment and cell genotype. As a result, the literature is filled with apparent contradictions of the effects of CTGF.
CTGF is a central mediator of tissue remodeling and fibrosis
CTGF has also been reported to inhibit BMP-7 signaling in diabetic kidney disease . Since BMP-7 is thought to counteract the pro-fibrotic effects of TGFβ [31, 32], inhibition of CTGF might also restore an anti-fibrotic regulatory pathway. Thus, inhibition of CTGF has the potential to modulate both pro- and anti-fibrotic mechanisms in a direction that could enable reversal of fibrotic processes.
In order to identify an inhibitor of CTGF with the potential to be developed into a therapeutic agent, FibroGen screened a library of human anti-CTGF antibodies to select those that bound to CTGF with reasonable affinity and exhibited activity in both in vitro assays and in vivo models of disease. FG-3019 was chosen as a clinical candidate that has now progressed to phase II testing. FG-3019 binds to the second domain of CTGF of all species tested. We have used FG-3019 in animal models of disease to test whether inhibition of CTGF could reverse pathophysiologic tissue remodeling and fibrosis.
CTGF is required for the fibrotic activity of TGFβ
Inhibition of CTGF can reverse fibrosis
Data from experiments in several organ systems suggest that inhibition of CTGF can not only prevent, but can also inhibit fibrosis. For example, in liver fibrosis models CTGF is known to increase in response to toxins such as CCl4  or dimethyl nitrosamine  or upon bile duct ligation . Inhibition of CTGF expression in a rodent model of CCl4-induced liver fibrosis with siRNA can prevent development of fibrosis . If the CTGF siRNA is administered to mice after significant increases of α-smooth muscle actin (αSMA, a marker of myofibroblasts) abundance and collagen deposition can be detected in their liver, the fibrosis appears to regress within two weeks .
To examine the cardiovascular complications of diabetes, rats were made diabetic with a single dose of streptozotocin. After 6 weeks of diabetes, the rats were divided into groups and treated for 6 weeks with FG-3019 (10 mg/kg tiw) or the angiotensin converting enzyme inhibitor captopril (75 mg/kg qd). The function of the hearts of the rats in the various groups was then measured prior to their sacrifice and removal of their carotid arteries for measurement of vascular stiffness. After 6 weeks of diabetes, the stiffness of the carotid arteries in the axial orientation was greater than that of healthy control animals. The axial stiffness continued to increase during the next 6 weeks of diabetes. In rats that began receiving captopril after 6 weeks of diabetes, further progression of vascular stiffening was halted, and the axial stiffness remained unchanged from that observed at 6 weeks. In contrast, in rats that received FG-3019 for 6 weeks beginning 6 weeks after they became diabetic, the axial stiffness of the carotid arteries in these diabetic rats reverted to that of healthy control animals. In addition the function of the heart could also be shown to be normalized in animals that received FG-3019, but not in those administered captopril . These data indicate that cardiovascular remodeling associated with the complications of diabetes can be reversed upon treatment with FG-3019.
To evaluate the effect of FG-3019 in pulmonary fibrosis, a radiation-induced model was used and preliminary data were reported at an American Thoracic Society meeting [39–41]. Pulmonary fibrosis was initiated with a single, full thorax irradiation (20 Gy) to mice and FG-3019 was administered for 8 weeks beginning at various times before or after irradiation. Lung density of all surviving mice was monitored by computed tomography (CT).
Irradiation induced lung remodeling beginning around 12 weeks, and FG-3019 attenuated this remodeling in a schedule-dependent manner. Lung density in the group administered FG-3019 beginning 16 weeks after irradiation had already significantly increased at the time that FG-3019 administration began, and decreased with treatment, indicating reversal of the radiation-induced changes .
Thus, inhibition of CTGF in liver, the cardiovascular system or the lungs has the potential to reverse tissue remodeling and the process of fibrosis. Together, the results from these studies suggest that inhibition of CTGF may benefit any disease in which tissue remodeling is important. For example, pancreatic cancers are known to be very desmoplastic  and to exhibit strong CTGF expression [43, 44]. Data suggest that CTGF can have both direct and indirect effects on pancreatic cancers [45–48]. Therefore, the use of FG-3019 for treatment of pancreatic cancer is being explored in parallel with hepatitis B-induced liver fibrosis and idiopathic pulmonary fibrosis.
This article has been published as part of Fibrogenesis & Tissue Repair Volume 5 Supplement 1, 2012: Proceedings of Fibroproliferative disorders: from biochemical analysis to targeted therapies. The full contents of the supplement are available online at http://www.fibrogenesis.com/supplements/5/S1.
We would like to thank our many colleagues at FibroGen and academic collaborators that have made contributions to the data discussed. In particular, we'd like to recognize Sebastian Bickelhaupt, Peter Huber and Peter Peschke of the DKFZ in Heidelberg, Germany for the data from the radiation-induced pulmonary fibrosis model and Chris Jacob and Ingrid Langsetmo of FibroGen for the data from the diabetic complications model. We'd also like to thank Carlita Fuentes for assistance in expediting preparation of this manuscript.
- De Winter P, Leoni P, Abraham D: Connective tissue growth factor: Structure-function relationships of a mosaic, multifunctional protein. Growth Factors. 2008, 26: 80-91. 10.1080/08977190802025602.View ArticlePubMedGoogle Scholar
- Holbourn KP, Acharya KR, Perbal B: The CCN family of proteins: structure-function relationships. Trends Biochem Sci. 2008, 33: 461-473. 10.1016/j.tibs.2008.07.006.PubMed CentralView ArticlePubMedGoogle Scholar
- Jaffa AA, Usinger WR, McHenry MB, Jaffa MA, Lipstiz SR, Lackland D: Connective tissue growth factor and susceptibility to renal and vascular disease risk in type 1 diabetes. J Clin Endocrinol Metab. 2008, 93: 1893-1900. 10.1210/jc.2007-2544.PubMed CentralView ArticlePubMedGoogle Scholar
- Nguyen TQ, Tarnow L, Andersen S, Hovind P, Parving HH, Goldschmeding R: Urinary connective tissue growth factor excretion correlates with clinical markers of renal disease in a large population of type 1 diabetic patients with diabetic nephropathy. Diabetes Care. 2006, 29: 83-88. 10.2337/diacare.29.01.06.dc05-1670.View ArticlePubMedGoogle Scholar
- Gressner AM, Yagmur E, Lahme B, Gressner O, Stanzel S: Connective tissue growth factor in serum as a new candidate test for assessment of hepatic fibrosis. Clin Chem. 2006, 52: 1815-1817. 10.1373/clinchem.2006.070466.View ArticlePubMedGoogle Scholar
- Kovalenko E, Tacke F, Gressner OA, Zimmermann HW, Lahme B, Janetzko A: Validation of connective tissue growth factor (CTGF/CCN2) and its gene polymorphisms as noninvasive biomarkers for the assessment of liver fibrosis. J Viral Hepat. 2009, 16: 612-620. 10.1111/j.1365-2893.2009.01110.x.View ArticlePubMedGoogle Scholar
- Bradham DM, Igarashi A, Potter RL, Grotendorst GR: Connective tissue growth factor: a cysteine-rich mitogen secreted by human vascular endothelial cells is related to the SRC-induced immediate early gene product CEF-10. J Cell Biol. 1991, 114: 1285-1294. 10.1083/jcb.114.6.1285.View ArticlePubMedGoogle Scholar
- Bornstein P, Sage E: Matricellular proteins: extracellular modulators of cell function. Curr Opin Cell Biol. 2002, 14: 608-616. 10.1016/S0955-0674(02)00361-7.View ArticlePubMedGoogle Scholar
- Leask A, Abraham DJ: All in the CCN family: essential matricellular signaling modulators emerge from the bunker. J Cell Sci. 2006, 119: 4803-4810. 10.1242/jcs.03270.View ArticlePubMedGoogle Scholar
- Shi-wen X, Leask A, Abraham D: Regulation and function of connective tissue growth factor/CCN2 in tissue repair, scarring and fibrosis. Cytokine Growth Factor Rev. 2008, 19: 133-144. 10.1016/j.cytogfr.2008.01.002.View ArticlePubMedGoogle Scholar
- Chen CC, Lau LF: Functions and mechanisms of action of CCN matricellular proteins. Int J Biochem Cell Biol. 2009, 41: 771-783. 10.1016/j.biocel.2008.07.025.PubMed CentralView ArticlePubMedGoogle Scholar
- Abreu JG, Ketpura NI, Reversade B, De Robertis EM: Connective-tissue growth factor (CTGF) modulates cell signalling by BMP and TGF-beta. Nat Cell Biol. 2002, 4: 599-604.PubMed CentralPubMedGoogle Scholar
- Inoki I, Shiomi T, Hashimoto G, Enomoto H, Nakamura H, Makino K: Connective tissue growth factor binds vascular endothelial growth factor (VEGF) and inhibits VEGF-induced angiogenesis. FASEB J. 2002, 16: 219-221.PubMedGoogle Scholar
- Nishida T, Kubota S, Fukunaga T, Kondo S, Yosimichi G, Nakanishi T: CTGF/Hcs24, hypertrophic chondrocyte-specific gene product, interacts with perlecan in regulating the proliferation and differentiation of chondrocytes. J Cell Physiol. 2003, 196: 265-275. 10.1002/jcp.10277.View ArticlePubMedGoogle Scholar
- Chen Y, Abraham DJ, Shi-wen X, Pearson JD, Black CM, Lyons KM: CCN2 (connective tissue growth factor) promotes fibroblast adhesion to fibronectin. Mol Biol Cell. 2004, 15: 5635-5646. 10.1091/mbc.E04-06-0490.PubMed CentralView ArticlePubMedGoogle Scholar
- Babic AM, Chen CC, Lau LF: Fisp12/mouse connective tissue growth factor mediates endothelial cell adhesion and migration through integrin alphavbeta3, promotes endothelial cell survival, and induces angiogenesis in vivo. Mol Cell Biol. 1999, 19: 2958-2966.PubMed CentralView ArticlePubMedGoogle Scholar
- Segarini PR, Nesbitt JE, Li D, Hays LG, Yates JR, Carmichael DF: The low density lipoprotein receptor-related protein/alpha2-macroglobulin receptor is a receptor for connective tissue growth factor. J Biol Chem. 2001, 276: 40659-40667. 10.1074/jbc.M105180200.View ArticlePubMedGoogle Scholar
- Wahab NA, Weston BS, Mason RM: Connective tissue growth factor CCN2 interacts with and activates the tyrosine kinase receptor TrkA. J Am Soc Nephrol. 2005, 16: 340-351. 10.1681/ASN.2003100905.View ArticlePubMedGoogle Scholar
- Wahab NA, Brinkman H, Mason RM: Uptake and intracellular transport of the connective tissue growth factor: a potential mode of action. Biochem J. 2001, 359: 89-97. 10.1042/0264-6021:3590089.PubMed CentralView ArticlePubMedGoogle Scholar
- Oliver N, Sternlicht M, Gerritsen K, Goldschmeding R: Could aging human skin use a connective tissue growth factor boost to increase collagen content?. J Invest Dermatol. 2010, 130: 338-341. 10.1038/jid.2009.331.View ArticlePubMedGoogle Scholar
- Gore-Hyer E, Shegogue D, Markiewicz M, Lo S, Hazen-Martin D, Greene EL: TGF-beta and CTGF have overlapping and distinct fibrogenic effects on human renal cells. Am J Physiol Renal Physiol. 2002, 283: F707-F716.View ArticlePubMedGoogle Scholar
- Paradis V, Dargere D, Bonvoust F, Vidaud M, Segarini P, Bedossa P: Effects and regulation of connective tissue growth factor on hepatic stellate cells. Lab Invest. 2002, 82: 767-774.View ArticlePubMedGoogle Scholar
- Grotendorst GR, Rahmanie H, Duncan MR: Combinatorial signaling pathways determine fibroblast proliferation and myofibroblast differentiation. FASEB J. 2004, 18: 469-479. 10.1096/fj.03-0699com.View ArticlePubMedGoogle Scholar
- Lee CH, Shah B, Moioli EK, Mao JJ: CTGF directs fibroblast differentiation from human mesenchymal stem/stromal cells and defines connective tissue healing in a rodent injury model. J Clin Invest. 2010, 120: 3340-3349. 10.1172/JCI43230.PubMed CentralView ArticlePubMedGoogle Scholar
- Hishikawa K, Oemar BS, Nakaki T: Static pressure regulates connective tissue growth factor expression in human mesangial cells. J Biol Chem. 2001, 276: 16797-16803. 10.1074/jbc.M010722200.View ArticlePubMedGoogle Scholar
- de Las HN, Ruiz-Ortega M, Miana M, Ruperez M, Sanz-Rosa D, Aragoncillo P: Interactions between aldosterone and connective tissue growth factor in vascular and renal damage in spontaneously hypertensive rats. J Hypertens. 2007, 25: 629-638. 10.1097/HJH.0b013e3280112ce5.View ArticleGoogle Scholar
- Chaqour B, Goppelt-Struebe M: Mechanical regulation of the Cyr61/CCN1 and CTGF/CCN2 proteins. FEBS J. 2006, 273: 3639-3649. 10.1111/j.1742-4658.2006.05360.x.View ArticlePubMedGoogle Scholar
- Yang H, Huang Y, Chen X, Liu J, Lu Y, Bu L: The role of CTGF in the diabetic rat retina and its relationship with VEGF and TGF-beta, elucidated by treatment with CTGFsiRNA. Acta Ophthalmol. 2010, 88: 652-659. 10.1111/j.1755-3768.2009.01641.x.View ArticlePubMedGoogle Scholar
- Liu FY, Xiao L, Peng YM, Duan SB, Liu H, Liu YH: Inhibition effect of small interfering RNA of connective tissue growth factor on the expression of vascular endothelial growth factor and connective tissue growth factor in cultured human peritoneal mesothelial cells. Chin Med J (Engl). 2007, 120: 231-236.Google Scholar
- Nguyen TQ, Roestenberg P, Van Nieuwenhoven FA, Bovenschen N, Li Z, Xu L: CTGF inhibits BMP-7 signaling in diabetic nephropathy. J Am Soc Nephrol. 2008, 19: 2098-2107. 10.1681/ASN.2007111261.PubMed CentralView ArticlePubMedGoogle Scholar
- Wahab NA, Mason RM: A critical look at growth factors and epithelial-to-mesenchymal transition in the adult kidney. Interrelationships between growth factors that regulate EMT in the adult kidney. Nephron Exp Nephrol. 2006, 104: e129-e134. 10.1159/000094963.View ArticlePubMedGoogle Scholar
- Mitu G, Hirschberg R: Bone morphogenetic protein-7 (BMP7) in chronic kidney disease. Front Biosci. 2008, 13: 4726-4739.View ArticlePubMedGoogle Scholar
- Mori T, Kawara S, Shinozaki M, Hayashi N, Kakinuma T, Igarashi A: Role and interaction of connective tissue growth factor with transforming growth factor-beta in persistent fibrosis: A mouse fibrosis model. J Cell Physiol. 1999, 181: 153-159. 10.1002/(SICI)1097-4652(199910)181:1<153::AID-JCP16>3.0.CO;2-K.View ArticlePubMedGoogle Scholar
- Paradis V, Dargere D, Vidaud M, De GA, Huet S, Martinez V: Expression of connective tissue growth factor in experimental rat and human liver fibrosis. Hepatology. 1999, 30: 968-976. 10.1002/hep.510300425.View ArticlePubMedGoogle Scholar
- Hsu YC, Chiu YT, Lee CY, Lin YL, Huang YT: Increases in fibrosis-related gene transcripts in livers of dimethylnitrosamine-intoxicated rats. J Biomed Sci. 2004, 11: 408-417. 10.1007/BF02254446.View ArticlePubMedGoogle Scholar
- Li G, Xie Q, Shi Y, Li D, Zhang M, Jiang S: Inhibition of connective tissue growth factor by siRNA prevents liver fibrosis in rats. J Gene Med. 2006, 8: 889-900. 10.1002/jgm.894.View ArticlePubMedGoogle Scholar
- Brigstock DR: Strategies for blocking the fibrogenic actions of connective tissue growth factor (CCN2): From pharmacological inhibition in vitro to targeted siRNA therapy in vivo. J Cell Commun Signal. 2009, 3: 5-18. 10.1007/s12079-009-0043-9.PubMed CentralView ArticlePubMedGoogle Scholar
- Langsetmo I, Jacob CT, Zhang W, Oliver N, Lin A, Coker G: Anti-CTGF human antibody therapy with FG-3019 prevents and reverses diabetes-induced cardiovascular complications in streptozotocin (STZ) treated rats. Diabetes. 2006, 55 (Suppl 1): A122.Google Scholar
- Huber PE, Bickelhaupt S, Peschke P, Tietz A, Wirkner U, Lipson KE: Reversal Of Established Fibrosis By Treatment With The Anti-CTGF Monoclonal Antibody FG-3019 In A Murine Model Of Radiation-Induced Pulmonary Fibrosis. Am J Respir Crit Care Med. 2010, 181: A1054.Google Scholar
- Plathow C, Li M, Gong P, Zieher H, Kiessling F, Peschke P: Computed tomography monitoring of radiation-induced lung fibrosis in mice. Invest Radiol. 2004, 39: 600-609. 10.1097/01.rli.0000138134.89050.a5.View ArticlePubMedGoogle Scholar
- Abdollahi A, Li M, Ping G, Plathow C, Domhan S, Kiessling F: Inhibition of platelet-derived growth factor signaling attenuates pulmonary fibrosis. J Exp Med. 2005, 201: 925-935. 10.1084/jem.20041393.PubMed CentralView ArticlePubMedGoogle Scholar
- Armstrong T, Packham G, Murphy LB, Bateman AC, Conti JA, Fine DR: Type I collagen promotes the malignant phenotype of pancreatic ductal adenocarcinoma. Clin Cancer Res. 2004, 10: 7427-7437. 10.1158/1078-0432.CCR-03-0825.View ArticlePubMedGoogle Scholar
- Wenger C, Ellenrieder V, Alber B, Lacher U, Menke A, Hameister H: Expression and differential regulation of connective tissue growth factor in pancreatic cancer cells. Oncogene. 1999, 18: 1073-1080. 10.1038/sj.onc.1202395.View ArticlePubMedGoogle Scholar
- Hartel M, di Mola FF, Gardini A, Zimmermann A, Di Sebastiano P, Guweidhi A: Desmoplastic reaction influences pancreatic cancer growth behavior. World J Surg. 2004, 28: 818-825. 10.1007/s00268-004-7147-4.View ArticlePubMedGoogle Scholar
- Karger A, Fitzner B, Brock P, Sparmann G, Emmrich J, Liebe S: Molecular insights into connective tissue growth factor action in rat pancreatic stellate cells. Cell Signal. 2008, 20: 1865-1872. 10.1016/j.cellsig.2008.06.016.View ArticlePubMedGoogle Scholar
- Dornhofer N, Spong S, Bennewith K, Salim A, Klaus S, Kambham N: Connective tissue growth factor-specific monoclonal antibody therapy inhibits pancreatic tumor growth and metastasis. Cancer Res. 2006, 66: 5816-5827. 10.1158/0008-5472.CAN-06-0081.View ArticlePubMedGoogle Scholar
- Bennewith KL, Huang X, Ham CM, Graves EE, Erler JT, Kambham N: The role of tumor cell-derived connective tissue growth factor (CTGF/CCN2) in pancreatic tumor growth. Cancer Res. 2009, 69: 775-784. 10.1158/0008-5472.CAN-08-0987.PubMed CentralView ArticlePubMedGoogle Scholar
- Aikawa T, Gunn J, Spong SM, Klaus SJ, Korc M: Connective tissue growth factor-specific antibody attenuates tumor growth, metastasis, and angiogenesis in an orthotopic mouse model of pancreatic cancer. Mol Cancer Ther. 2006, 5: 1108-1116. 10.1158/1535-7163.MCT-05-0516.View ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.